犬肾细胞培养流感病毒条件的优化

目的 对影响流感病毒细胞分离培养的因素进行优化,得到细胞分离培养流感病毒的最佳方案。方法 将4种型别流感毒株接种至犬肾上皮细胞（MDCK）,以不同接种量、吸附时间、培养时间以及TPCK胰蛋白酶浓度进行病毒分离培养,以流感病毒红细胞凝集实验的血凝滴度作为指标对流感分离培养结果进行评价分析。结果甲型H1N1流感病毒、季节性流感病毒H3N2、乙型Victoria系流感病毒、乙型Yamagata系流感病毒在培养过程中的最适条件分别为：接种量为200、200、250、300μL/孔,吸附时间为1.5、1.5、2.0、2.0 h,TPCK胰酶浓度为1.0、2.0、2.0、1.5 mg/mL,培养时间为96、72、120、120 h。结论 成功对4种亚型流感病毒在MDCK细胞中的培养条件进行了优化,在培养流感病毒时应按照不同型别选取不同的培养条件。     

流感病毒在不同细胞中适宜培养条件的研究

目的 探索流感病毒在细胞中培养的适宜条件.方法 采用MEK细胞、FRhK-4细胞、MDCK细胞分别接种甲型流感病毒H1N1型、H3N2型和乙型流感病毒疫苗毒株,观察3种毒株在不同培养条件下的细胞病变、血凝滴度.结果 3株流感病毒接种细胞在33℃培养72 h后出现拉网、圆缩、脱落等现象;FRhK-4细胞和MDCK细胞对流感病毒较敏感.胞龄为48 h的细胞,在NaHCO3的终浓度为1.4%o,TPCK-胰酶浓度为1.0μg/ml的病毒维持液培养流感病毒为最适条件.结论 寻找到了细胞流感病毒的适宜条件.     

新复苏MDCK细胞培养甲型H3N2流感病毒方法的优化

目的优化甲型H3N2流感病毒在新复苏的犬肾细胞(MDCK)上的培养条件,提高该病毒在新复苏细胞上培养的分离效果。方法选取6例复核鉴定过的甲型H3N2病毒株,以吸附法分别接种于新复苏的MDCK细胞上培养,检测收获病毒液HA滴度,通过比较不同的细胞胞龄、细胞代次、病毒接种吸附时间对甲型H3N2流感病毒易感性的影响,确定最佳培养条件。结果不同胞龄MDCK细胞接种H3N2病毒后,胞龄为3 d、4 d的细胞所收获的病毒液HA滴度均明显高于其他组,差异有统计学意义(F=1. 279,P=0. 027);对复苏后第2代、第3代、第4代细胞进行接种后,H3N2病毒HA均明显高于第一代细胞,差异有统计学意义(F=0. 765,P=0. 008);病毒接种吸附时间(1～2 h)对甲型H3N2病毒HA滴度没有明显影响,差异无统计学意义(F=2. 743,P=0. 178)。结论甲型H3N2病毒接种于新复苏MDCK细胞最佳培养条件为:选用复苏后传代培养至第二代及以后、胞龄3～4 d的MDCK细胞进行接种,接种吸附时间1～2 h之内,可获得HA滴度较高的甲型H3N2流感病毒液。     

MDCK、Vero和Hep-2细胞共培养分离甲型H1N1型流感病毒

目的用MDCK、Vero和Hep-2细胞共培养体系,分离甲型H1N1流感病毒,提高流感监测效率及缩短临床诊断周期。方法选取4株经国家流感中心复核鉴定、本实验室保存的甲型H1N1流感病毒,用病毒培养液1:2稀释后分别接种MDCK、Vero和Hep-2共培养细胞瓶和MDCK细胞瓶,每组设空白对照,用生理盐水代替接种。经35℃、5%CO2培养后记录甲型H1N1型流感病毒毒株在共培养细胞瓶和MDCK细胞瓶中的细胞病变效应(CPE)、血凝滴度和荧光定量PCR反应的CT值。结果培养后,2组接种流感病毒的细胞瓶均出现典型细胞病变,MDCK细胞瓶CPE略强于共培养细胞瓶,分离物-70℃冻融2次后检测血凝滴度,2组接种流感病毒的细胞瓶内分离物,血凝滴度均(1∶32;提取分离物核酸,荧光定量PCR检测,CT值均(20。结论用MD-CK、Vero和Hep-2细胞共培养体系可以分离流感病毒,同时还可分离鉴定其它呼吸道病毒。     

流感病毒在MDCK细胞中分离与培养的影响因素

目的探求流感病毒在MDCK（狗肾细胞）中培养的影响因素。方法将流感病毒吸附接种于MDCK细胞,对一些因素（样本处理情况、吸咐和培养时间、血清浓度、胰酶浓度、接种量和传代次数等）进行比较。结果样本反复挤压,最佳收获病毒时间为72h;加牛血清白蛋白组分V和维持液中终浓度为2．5μg/ml的胰酶可明显提高细胞增殖病毒的血凝效价。结论通过对MDCK细胞培养与增毒的影响因素分析,明显提高流感病毒分离与增毒的效价。     

单纯疱疹病毒Ⅱ型在Vero细胞中培养与增殖特性的研究

目的研究单纯疱疹病毒Ⅱ型（HSV-2）在非洲绿猴肾细胞（Vero细胞）中培养及增殖的合适条件。方法把HSV-2接种于Vero细胞中培养和传代,于不同时间收获病毒液并测毒力,进行不同培养条件的摸索。结果pH值、残留牛血清是HSV-2在Vero细胞培养的影响因素,最佳收毒时间为36-72h。在Vero细胞上培养病毒3-5代,病毒毒力较高。结论建立了单纯疱疹病毒Ⅱ型在Vero细胞上培养增殖的初步方法,为下一步建立HSV-2的潜伏与激发细胞模型打下基础。     

深圳南山区2005年流行性感冒病原学监测分析

目的了解流感病毒的流行情况，探讨流感流行规律，为制订流感控制措施提供依据。方法对辖区监测门诊及疫情点的疑似流感样病例采集咽拭子标本，接种狗肾传代细胞（MDCK）培养进行流感病毒分离及进行血凝试验，并对阳性者进行血凝抑制试验进行分型鉴定。结果全年检测流感样标本与咽拭子标本共480份，分离出流感病毒64株，分离率13．33％。分别为H1N1亚型9株、H3N2亚型44株、B型11株。结论2005年深圳市南山区是以H3N2亚型流感病毒流行为主，九月份以H3N1亚型和B型流感流行为主。     

上海2株严重急性呼吸综合征冠状病毒2的分离与鉴定

目的 从新型冠状病毒肺炎（COVID-19）患者鼻/咽拭子样本中分离、培养严重急性呼吸综合征冠状病毒2（SARS-CoV-2）。方法 将来自上海市COVID-19患者的3份鼻/咽拭子样本以TPCK胰酶处理,然后接种Vero E6细胞;待大部分细胞出现明显病变时,取细胞培养上清用qRT-PCR法检测病毒核酸,并用反转录PCR扩增病毒受体结合区（RBD）基因片段;将病毒扩增培养后感染接种于96孔板中的Vero E6细胞,观察细胞病变效应,并用免疫荧光法检测病毒蛋白。结果 2份COVID-19患者鼻/咽拭子样本接种的Vero E6细胞出现明显细胞病变效应,细胞培养上清中检测出新复制产生的SARS-CoV-2核酸,扩增出的RBD序列与早期分离出的SARS-CoV-2相应序列完全一致;病毒感染的Vero E6细胞病变迅速,并能与SARS-CoV-2核衣壳蛋白（N蛋白）单克隆抗体、刺突蛋白（S蛋白）单克隆抗体及COVID-19患者恢复期血清发生反应。结论 从2份COVID-19患者鼻/咽拭子样本中成功分离出SARS-CoV-2,为后续开展SARS-CoV-2感染与致病机制研究、防治药物与疫苗的研发奠定了基础。     

应用生物反应器培养Vero细胞制备甲肝病毒

目的　研究用生物反应器培养Vero细胞制备甲肝病毒W株的可行性。方法　Vero细胞悬浮吸附HAVW株后 ,接种到搅拌式生物反应器中 ,用微载体进行培养 ,对营养物质、代谢产物及病毒的增殖进行分析。结果　经四周的培养 ,病毒收获时 ,细胞密度达 1.4× 10 6 ml,甲肝病毒抗原滴度达 1∶64。结论　W株已适应于Vero细胞中 ,用生物反应器培养产毒 ,有较好的应用前景     

2株广西鸭源H6N2亚型禽流感病毒遗传进化以及对BALB/c小鼠的致病性分析

目的：探讨广西鸭源H6N2亚型禽流感病毒的分子遗传特征，评估其对哺乳动物BALB/c小鼠的致病性。方法：选择前期在广西分离的2株鸭源性H6N2禽流感病毒A/DK/GX/3101/2018(GX3101)、A/DK/GX/5220/2018(GX5220)，基因测序和遗传进化分析病毒全基因组的核苷酸同源性及遗传进化，受体结合特异性法分析2株病毒的受体结合特性，应用BALB/c小鼠评估鸭源H6N2病毒的致病性。结果：2株H6N2病毒属于欧亚谱系，HA裂解位点处为PQIETG/GL，属低致病性禽流感病毒特征，HA蛋白氨基酸位点出现E190V位点的氨基酸突变，2株病毒均识别与结合禽型SAα-2,3 Gal受体。病毒接种小鼠后，肺组织病毒分离培养阳性，小鼠肺组织出现炎症细胞浸润、肺泡结构完整性破坏，免疫组化检测病毒抗原显示，GX3101病毒接种小鼠的肺组织出现流感病毒核蛋白（NP）阳性。结论：广西鸭源H6N2毒株具有禽流感病毒的特点，可不经适应直接跨种属间屏障感染哺乳动物小鼠，须密切监测家禽中流感病毒H6N2流行和进化演变。     

Value of D-Dimers in patients with acute aortic dissection

Objective: To evaluatel the value of D-dimers in patients with acute aortic dissection (AAD). Methods: This study consisted of 16 patients with AAD and 27 non-AAD patients. Serum D-dimets were measured by Sta-Liatest D-DI immunoturbidimetric assay. Results: D-dimer level was higher (P ＜ 0.001) in patients with AAD(7.91 ± 5.52 μg/ml) than that in non- AAD group(1.57±1.24 μg/ml). D-dimer was positive (＞0.4 μg/ml) in all patients with AAD and in 10 control group patients (37%). Among patients with acute AAD, D-dimers tended to be higher in Stanford A than in Stanford B (8.67 ± 4.31 μg/ml vs. 3.24±1.27 μg/ml, P ＜0.01). D-dimer values tended to be higher in more extended disease(3.84 ± 1.65 μg/ml, 8.57 ± 3.58 μg/ml and 11.87 ± 5.69 μg/ml in thoracic aorta, thoracic and abdominal aorta, thoracic and abdominal aorta and iliacal arteries, respectively, P ＜ 0.05 for both 8.57 ± 3.58 and 11.87 ± 5.69 vs. 3.84 ± 1.65 ). Including the control group into the analysis, we found a sensitivity of 100%, a negative predictive value of 100%, and a specificity of 66% and a positive predictive value of 64% for D-dimer in diagnosis of AAD in our patients with suspected AAD. Conclusion: D-dimer was elevated in patients with AAD. A negative D-dimer test result could be useful in excluding AAD.     

Research of combined liver-kidney transplantation model in rats

Objective： To set up a simple and reliable rat model of combined liver-kidney transplantation. Methods： SD rats served as both donors and recipients. 4℃ sodium lactate Ringer＇s was infused from portal veins to donated livers,and from abdominal aorta to donated kidneys, respectively. Anastomosis of the portal vein and the inferior vena cava （IVC） inferior to the right kidney between the graft and the recipient was performed by a double cuff method, then the superior hepatic vena cava with suture. A patch of donated renal artery was anastomosed to the recipient abdominal aorta. The urethra and bile duct were reconstructed with a simple inside bracket. Results： Among 65 cases of combined liver-kidney transplantation, the success rate in the late 40 cases was 77.5%. The function of the grafted liver and kidney remained normal. Conclusion： This rat model of combined liver-kidney transplantation can be established in common laboratory conditions with high success rate and meet the needs of renal transplantation experiment.     

APPLICATION OF LORNOXICAM TO PATIENT-CONTROLLED ANALGESIA IN PATIENTS UNDERGOING ABDOMINAL SURGERIES

FOR anesthesiologis s ,treatingpostoperativepainhas alwaysbeen a problem.Althoughopioidshave been provedtobe effective,theirsideeffectscouldnotbeignored.With thedevelopmentofscienceand pharmacology,many drugs with aspectsof satisfactoryanalgesicefficacyand couldbe welltoleratedby patientshave been developed.And lornoxicamisone of them, which isa non-steroidalanti-inflammatorydrug (NSAID ), with analgesic, anti-infl-ammatory,andantipyreticproperties.Itseliminationhalf-time(3 to 5 hours) isle…     

BLOOD PRESSURE CHANGE WITH AGE IN SALT-SENSITIVE TEENAGERS

Objective To observe blood pressure change with age in salt-sensitive teenagers whose salt sensitivity were determined by repeated testing.Methods Salt sensitivity was determined through intravenous infusion of normal saline combined with volume-depletion by oral diuretic furosemide in 55 teenagers. After five years, salt sensitivity was re-examined and subject blood pressure was followed up. Blood pressure changes in salt-sensitive teenagers were compared to that of non-salt sensitive teenagers over five years.Results After 5 years, the repetition rate of salt sensitivity determined by intravenous saline loading is 92.7%. In teenagers with salt sensitivity on the baseline, both the systolic blood pressure increments and increment rates were much higher than non-salt sensitive teenagers (12.7±12.1 mmHg vs. 2.8±5.2 mmHg, P＜ 0.01; 12.2%± 12.0% vs. 2.5% ±4.4%, P＜ 0.001,respectively). There was a similar trend for diastolic blood pressure (8.4 ± 6.4 mmHg vs. 3.7 ± 6.4 mmHg, P = 0.052; 13.2% ±10.6 % vs. 6.8%± 10.1%, P = 0.053, respectively).Conclusions Salt sensitivity determined by intravenous saline loading showed good reproducibility. Blood pressure increments with age were much higher in salt-sensitive teenagers than non-salt sensitive teenagers, especially in terms of systolic blood pressure.     

Development on Adjustable Calibration Marker for Shock Wave Focus

Shock wave lithotripsy （SWL） is a treatment of choice for upper urinary stones. However, this procedure is inappropriate for obese patients because the focus is often unable to reach the target owing to the limited focal distance in shock wave source. Although treating such patients in a blast path may increase the application length of shock wave source, it＇s difficult to find this path on the lithotripter monitor. For this reason, we invented an adjustable calibration marker in order to set an effective focus in the shock wave hath.     

Study on the TCM Pathogenesis of Angiopathic Complications of Type II Diabetes

Excess production of reactive oxygen species(ROS)of mitochondrion mediated by hyperglycemia is the common pathogenesis of angiopathic complications of diabetes.TCM holds that the damp from the dysfunction of spleen.kidney and liver is the causative factor of complications of diabetes.This is similar to the mechanism of Ros resulting in angiopathic complications of diabetes.When the angiopathic complications of type II diabetes mellitus(T2DM)are difierentiated as caused by turbid damp in TCM can be explained as ROS.Since the obstruction of pathogenic damp in channels and collaterals is said to be the main pathogenesis,the treating principle should be dissolving the damp to remove the obstruction.     

Leptin, Ghrelin and TNF-α before and after Hypo-caloric Traditional Chinese Diet and Auricular Acupuncture

INTRODUCTION Obesity is a complex emergent problem, which can be possibly solved not only by the diet but also by the life style and promotion of a constant physical exercise. 1, 2 No doubt careful attentions must be given to the nutritional condition of obese people, the dietary habits, the somatic build (i.e. distribution of fat mass) and the organic functions linked to formation of the fat mass. All the parameters should be constantly monitored before, during and after a diet treatment. 3, 4, 5     

Relationship between Dysglycemia and Carotid Atherosclerosis in Tibetan Population

People with dysglycemia are at high risk for atherosclerotic diseases. This study aims at investigating the atherosclerotic vascular damage in dysglycemia and its metabolic origin in Tibetan population.     

Acupuncture Combined with Spinal Tui Na for Treatment of Primary Dysmenorrhea in 30 Cases

Objective: To observe the therapeutic effects in acupunture treatment of primary dysmenorrhea combined with spinal Tui Na, and study its mechanism. Methods: Thirty cases of the treatment group were treated by acupuncture combined with spinal Tui Na, and thirty cases in the control group were treated by routine acupuncture. Results: The total effective rate was 93.3% in the treatment group, and 73.3% in the control group, with a significant difference between the two groups (P<0.05). Conclusions: Acupuncture combined with spinal Tui Na has good prospects for treatment of primary dysmenorrhea.     

Luo Lingjie's Experience in Treating Chronic Nephropathy

In treating chronic nephropathy,Luo Lingjie,a chief physician,pays attention to regulating the balance between yin and yang,treating infection if present,and removing pathogenic factors.He prescribes gentle drugs and uses carefully strongly warming-tonifying ones,emphasizes the importance of persuading the patient to persist in treatment with medication and nurse one's health for recuperation,and is good at combined use of TCM and western medicine therapy and brings the merits of various therapies into full play,with obvious theraoeutic effects.